IZSTO Istituto Zooprofilattico Sperimentale del Piemonte, Liguria e Valle d’Aosta Entomological surveillance and Public Health: first detection of West Nile virus in Piemonte and Liguria and Aedes albopictus in Valle d'Aosta (North western Italy) Verna F.1 , Francese D.R.1, Pautasso A 1, Radaelli M.C.1, Mosca A.2, Grattarola C.1, Desiato R.1, Bertolini S.1, Lia R.1, Monaci E.1, Vitale, N.1 , Modesto P.1, Ballardini M.1 , Orusa R.1, Spedicato R.1, Accorsi A.1, Mignone W.1, Prearo M.1, Chiavacci L.1, Casalone C.1 1 Istituto Zooprofilattico del Piemonte, Liguria e Valle d’Aosta, Torino, Italy (www.izsto.it; [email protected]) 2 Istituto Per le Piante da Legno e l’Ambiente, Torino, Italy Indroduction Mosquito Borne Diseases (MBDs) as West Nile and Usutu represents an emerging threat for both humans and animals. Since 2011 Istituto Zooprofilattico Sperimentale of Piemonte, Liguria e Valle d’Aosta (IZS PLVA), through research projects funded by the Ministry of Health and European Regional Development Fund, has been implementing entomological surveillance for West Nile Virus (WNV) in its territories (North-western Italy), considered WNV free, in addition to the activities supported by the Ministry of Health at national level and mainly focused in endemic areas. In this work we report results of entomological surveillance carried out in 2014 in Piemonte, Liguria and Valle d'Aosta Regions. Materials and methods Fig 1: collection sites According to risk factors 46 collection sites (respectively 34 in Piemonte, 6 in Liguria and 6 Valle d’Aosta regions) were selected (Fig 1). From July to November, mosquitoes were fortnightly trapped using CO2 CDC dry ice-baited traps, BG-Sentinel traps and Gravid traps (Fig. 2). In order to detect the introduction of exotic mosquitoes, ovitraps were also placed in airports and ports. Eggs were incubated and emerged adults were identified. Adult mosquitoes collected were identified to species level by morphological standard classification keys, pooled and analyzed by Real Time RT-PCR distinctive for WNV Lineage 1 and 2 and Usutu virus (USUV). Results A total of 20036 adult mosquitoes belonging to 16 different species were collected and 916 pools tested (Fig. 3). WNV Lineage 2 was detected in 3 pools of Culex pipiens: 2 in Piemonte (Alessandria province) and 1 in Liguria (Genoa). This is the first report of WNV circulation in both regions. Moreover USUV was detected in two pools of Culex pipiens, collected in Piemonte (Alessandria) and Liguria (La Spezia). Control measures against WNV and its vector were quickly implemented, coordinated by IZS PLVA and together with veterinary and human local health units, as provided by national legislation (screening of equine blood samples and blood transfusion) and by local protocols applied by Istituto Per le Piante da Legno e l’Ambiente (adulticiding and larviciding). Moreover, monitoring activities were also implemented by identifying further 10 sites. From ovitraps, 677 mosquitoes emerged, identified as Aedes albopictus (respectively 26 in Valle d'Aosta, 31 in Piedmont and 620 in Liguria). By means of ovitraps, for the first time Aedes albopictus was identified in Valle d’Aosta, previously free. Other species: Culex modestus Culiseta annulata Anopheles plumbeus Culex territans Culiseta subhocrea Culex hortensis Culex theileri Ochlerotatus geniculatus Piemonte Culex impudicus Liguria Culex mimeticus Valle d’Aosta Culiseta longiareolata Fig 4: mosquitoes species trapped in the three Regions in 2014. Conclusion a b c d Other Aedes vexans Anopheles maculipennis Ochlerotatus caspius Aedes albopictus 8000 7000 6000 5000 4000 3000 2000 1000 0 Culex pipiens 9000 Fig.2: a) G-Sentinel, b) Gravid trap, c) CDC trap, d) ovitrap Fig 4: traps location where USUV and WNV were detected Our research projects show that the entomological surveillance is necessary to early detect the introduction of exotic mosquitoes and the presence of WNV before the appearance of clinical cases in both humans and animals. It allows to quickly implement control measures aiming to protect human health, through veterinary and human Authorities. USUV circulation, reported in our territories since 2011, should be more considered given the report of recent neuroinvasive human cases in Croatia. References Del Amo J, Sotelo E, Fernández-Pinero J, Gallardo C, Llorente F, Agüero M, Jiménez-Clavero MA. A novel quantitative multiplex real-time RT-PCR for the simultaneous detection and differentiation of West Nile virus lineages 1 and 2,and of Usutu virus. J Virol Methods. 189(2):321-7. 2013 Santini M1, Vilibic-Cavlek T, Barsic B, Barbic L, Savic V, Stevanovic V, Listes E, Di Gennaro A, Savini G. First cases of human Usutu virus neuroinvasive infection in Croatia, August-September 2013: clinical and laboratory features. J Neurovirol. 2015 Feb;21(1):92-7. Aknowledgments This study was supported by a grant from the European Regional Development Fund (REDLAV) and a grant from Italian Ministry of Health, “Ricerca corrente 2011 (11RC07)”. GERI2015 – GENES, ECOSYSTEMS AND RISK OF INFECTION. Heraklion 21-23 April 2015.